CN108865930A - RADIX CURCUMAE endogenetic bacteria bacterial strain ZJU-C612-2 and its application - Google Patents
RADIX CURCUMAE endogenetic bacteria bacterial strain ZJU-C612-2 and its application Download PDFInfo
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Abstract
The invention discloses a kind of Bei Laisi bacillus velezensis ZJU-C612-2, deposit numbers：CCTCC NO：M 2018312.Bei Laisi bacillus velezensis ZJU-C612-2 can inhibit the growth of dendrobium candidum anthrax bacteria, Crocus bulb rotten pathogenic bacteria, walnut branch-rot bacterium, Sclerotinia sclerotiorum, cotton standing dead silk core germ, Didymella bryoniae；Dendrobium candidum protocorm and tissue-cultured seedling can also be promoted to grow.
Description
Technical field
The invention belongs to microorganism field, antibacterial, growth-promoting functions the RADIX CURCUMAE endogenetic bacteria ZJU-C612- of specially one plant tool
2 and its application.
Background technique
Endophyte of plant (Endophyte) refers to that a certain stage in the full history of life or the history of life parasitizes plant tissue
In, and the microorganism for not causing clearly visible disease and function to change to plant itself.Studies have shown that many endophyte of plant tools
There are biological and ecological methods to prevent plant disease, pests, and erosion and growth-promoting functions.Endophyte Biocontrol Effect mechanism mainly include nutrition and space site competition, generate antibacterial substance and
Induce disease resistance of plant.And, mainly there are two aspects in the reason of endophyte generation growth-promoting functions, on the one hand, many endophytes can produce
Auxins, these hormones such as raw auxin, gibberellin and cytokinin can effectively facilitate the growth and development of plant；
On the other hand, endophyte can enhance the nitrogen fixing capacity of plant and the absorbability to nutrients such as potassium, calcium, phosphorus, iron.
RADIX CURCUMAE is Zingiber curcuma genus perennial herb, first recorded in《Tang materia medica》, it is distributed mainly on Wenzhou District of Zhejiang Province
Ruian is famous one of " eight Zhe's ".RADIX CURCUMAE endophyte resource rich in collects the report of RADIX CURCUMAE endophyte more
In, current separated acquisition 7 sections upper (Yan et al., 2014) in endogenetic fungus separation and the research of endophyte secondary metabolite
Totally 30 category endogenetic fungus, and the biological and ecological methods to prevent plant disease, pests, and erosion of RADIX CURCUMAE endogenetic bacteria research be rarely reported so that RADIX CURCUMAE endogenetic bacteria this
One valuable source is not fully developed, has biological and ecological methods to prevent plant disease, pests, and erosion, the R and D of growth-promoting potential endogenetic bacteria in RADIX CURCUMAE for this purpose, carrying out
It is of great significance.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of Bei Laisi bacillus with biological and ecological methods to prevent plant disease, pests, and erosion and growth-promoting functions
(Bacillus velezensis) new strains ZJU-C612-2 and application thereof.
In order to solve the above technical problem, the present invention provides Bei Laisi bacillus velezensis ZJU-
C612-2, preservation title：Bei Laisi bacillus ZJU-C612-2Bacillus velezensis ZJU-C612-2, preservation list
Position：China typical culture collection center, preservation address：Wuhan, China Wuhan University, deposit number：CCTCC NO：M
2018312, preservation date：On May 28th, 2018.
Bei Laisi bacillus velezensis ZJU-C612-2 is able to suppress dendrobium candidum anthrax bacteria
(Colletotrichum gloeosporioides), Crocus bulb rotten pathogenic bacteria (Fusarium oxysporum), walnut
Branch-rot bacterium (Botryosphaeria dothidea), Sclerotinia sclerotiorum (Sclerotinia sclerotiorum), cotton
6 kinds of plants of miliary damping-off germ (Rhizoctonia solani) and Didymella bryoniae (Mycosphaerella melonis)
The growth of pathogenic bacteria mycelia.
Bei Laisi bacillus velezensis ZJU-C612-2 can promote dendrobium candidum protocorm and
The growth of tissue-cultured seedling.
The present invention has found that this plant has the RADIX CURCUMAE endogenetic bacteria of biological and ecological methods to prevent plant disease, pests, and erosion and growth-promoting functions in RADIX CURCUMAE endogenetic bacteria
Bacterial strain ZJU-C612-2, the identified bacterium are Bei Laisi bacillus.It can generate a variety of biocidal property active materials, including egg
White enzyme, 1,4 beta-glucanase, thermophilic iron element and lipopeptide antibiotic, to dendrobium candidum anthrax bacteria, Crocus bulb rotten pathogenic bacteria, core
The growth of peach branch blight bacterium, 6 kinds of Sclerotinia sclerotiorum, cotton standing dead silk core germ and Didymella bryoniae plant pathogen mycelia
With good inhibiting effect.In addition, the bacterium can also generate gibberellin (GA), zeatin (ZT), kinetin (KT), auxin
(IAA) and 5 Plant Hormone of witchweed lactone (SL), there is facilitation to the growth of dendrobium candidum protocorm and tissue-cultured seedling.
The development and utilization for being found to be RADIX CURCUMAE endogenetic bacteria of the bacterium provides good basis.
Present invention finds the bacterial strain ZJU-C612-2 that one plant has biological and ecological methods to prevent plant disease, pests, and erosion, growth-promoting functions, which is isolated from RADIX CURCUMAE, is
RADIX CURCUMAE endogenetic bacteria.Although it is reported that RADIX CURCUMAE endophyte has Biocontrol Effect (Wang et al., 2012), this
A little endophytes are endogenetic fungus, rather than endogenetic bacteria.
Identified bacterial strain ZJU-C612-2 is Bei Laisi bacillus, which is able to suppress dendrobium candidum anthrax bacteria, west
Safflower stem-bulb rot germ, walnut branch-rot bacterium, Sclerotinia sclerotiorum, cotton standing dead silk core germ and 6 kinds of Didymella bryoniae
The growth of plant pathogen mycelia.Although informing Bei Laisi gemma bar in patent CN 102703342 and CN 102286412
Bacterium can be respectively used to prevention and treatment Hybrid Bamboo top dry and tomato wilt, but the pathogenic bacteria of both diseases and of the invention 6 kinds
Plant pathogen is different.In addition, there is document report Bei Laisi bacillus to can be used in preventing and treating wheat powdery mildew (Cai et
Al., 2017), the diseases such as black spot of cabbage (Shuna et al., 2011) and wheat scab (Juan et al., 2007),
These diseases are not also identical as 6 kinds of diseases of this research.
The fermentation liquid of bacterial strain ZJU-C612-2 can promote the growth of dendrobium candidum protocorm and tissue-cultured seedling.Although Meng Q
It can promote to some extent beet Deng (2016) discovery Bei Laisi bacillus BAC03, carrot, cucumber, pepper, potato, trailing plants
Foretell, pumpkin, the growth of totally 9 kinds of plants such as tomato and radish, but about Bei Laisi bacillus promote dendrobium candidum protocorm and
The document of the growth of tissue-cultured seedling has not been reported.
Detailed description of the invention
Specific embodiments of the present invention will be described in further detail with reference to the accompanying drawing.
Fig. 1 is bacterial strain ZJU-C612-2 screening experiment figure；A：Dendrobium candidum anthrax bacteria；B：Crocus bulb rot disease
Bacterium.
Fig. 2 is bacterial strain ZJU-C612-2 morphological feature figure；A：Colonial morphology of the bacterial strain ZJU-C612-2 on LB culture medium；
B：The thalli morphology of bacterial strain ZJU-C612-2 electricity microscopic observation.
Fig. 3 is that bacterial strain ZJU-C612-2 produces protease, chitinase, 1,4 beta-glucanase, thermophilic iron element ability measurement result figure；
A：Protease；B：Chitinase；C：1,4 beta-glucanase；D：Thermophilic iron element.
Fig. 4 is that bacterial strain ZJU-C612-2 produces lipopeptide antibiotic mass spectrogram；A：The mass spectrogram of m/z 800~3000；B：Matter
The partial enlargement of m/z 1000~1100 in spectrogram A；C：The partial enlargement of m/z 1400~1500 in mass spectrogram A.
Fig. 5 is the chromatogram of GA, KT, ZT, IAA and SL in bacterial strain ZJU-C612-2 fermentation extracting solution；
A1, A2, A3, A4, A5 are respectively the chromatogram of GA, KT, ZT, IAA and SL standard specimen；
B1, B2, B3, B4, B5 are respectively the chromatogram of GA, KT, ZT, IAA and SL in fermentation liquid.
Fig. 6 is antagonistic effect figure of the bacterial strain ZJU-C612-2 to 6 kinds of plant pathogens；A：Dendrobium candidum anthrax bacteria；B：
Crocus bulb rotten pathogenic bacteria；C：Walnut branch-rot bacterium；D：Sclerotinia sclerotiorum；E：Cotton standing dead silk core germ；F：Watermelon is climing
Blight bacterium.
Fig. 7 is growth-promoting functions figure of the bacterial strain ZJU-C612-2 to dendrobium candidum protocorm；A：Sterile LB control；B：ZJU-
C612-2 bacterial strain.
Fig. 8 is growth-promoting functions figure of the bacterial strain ZJU-C612-2 to candidum tissue culturing seedling；A：Sterile LB control；B：ZJU-
C612-2 bacterial strain.
Specific embodiment
The present invention is described further combined with specific embodiments below, but protection scope of the present invention is not limited in
This：
The screening of embodiment 1, bacterial strain ZJU-C612-2
Healthy RADIX CURCUMAE root tuber picks up from Lishui Yunhe County, and the separation of endogenetic bacteria uses conventional organization partition method
(it can refer to CN103122331A's《A kind of separation method of endophyte of plant》), it isolates and obtains 25 plants of endogenetic bacterias.
The screening of antagonistic strain uses tablet face-off method, comprises the concrete steps that：
(1) endogenetic bacteria is activated on LB solid plate, chooses the LB liquid medium that a ring single colonie is inoculated into 4ml
In, 37 DEG C are placed in, cultivates for 24 hours as strain, is inoculated with 1% (volume %) inoculum concentration interior raw thin in 200r/min shaken cultivation case
Bacterium is into new LB liquid medium, and 37 DEG C, 200r/min shaken cultivation for 24 hours, it is spare to obtain endogenetic bacteria fermentation liquid.
(2) using dendrobium candidum anthrax bacteria and Crocus bulb rotten pathogenic bacteria as indicator bacteria, the screening of Antagonistic Fungi is carried out.It takes
On dendrobium candidum anthrax bacteria and Crocus bulb rotten pathogenic bacteria inoculated by hypha block to new PDA culture medium, activated at 28 DEG C
7d is cultivated, takes the bacteria cake of diameter 5mm with punch along colony edge, is put into plate center, is set away from point at the 2.0cm of bacterium colony two sides
5ml endogenetic bacteria fermentation liquid observes each bacterial strain to the inhibitory effect of indicator bacteria, the antibacterial bandwidth in two sides is measured after 4d, and ask flat
Mean value.
It is equal to dendrobium candidum anthrax bacteria and Crocus bulb rotten pathogenic bacteria by one plant of the screening experiment acquisition of Antagonistic Fungi
Bacterial strain ZJU-C612-2 with preferable fungistatic effect, as shown in Figure 1.
Bacterial strain ZJU-C612-2 preservation, preservation title have been subjected to：Bei Laisi bacillus ZJU-C612-2Bacillus
Velezensis ZJU-C612-2, depositary institution：China typical culture collection center, preservation address：Wuhan, China Wuhan is big
It learns, deposit number：CCTCC NO：M 2018312, preservation date：On May 28th, 2018.
The identification of embodiment 2, bacterial strain ZJU-C612-2
Amplification sequencing is carried out with 16S rDNA gene order of the universal primer F27 and R1522 to bacterial strain ZJU-C612-2, is obtained
The sequence for being 1487bp to size, sequence is uploaded to and obtains gene accession number in GenBank：KY078349.1.By the sequence
Blast is carried out on the website NCBI and compares analysis, as the result is shown itself and Bei Laisi bacillus (CP023414.1), solution starch bud
The similarity of spore bacillus (KT961125.1) and bacillus subtilis (HQ711983.1) is 99%.
After bacterial strain ZJU-C612-2 is cultivated for 24 hours on LB plate, the single colonie of formation is similar round, and edge is irregular, has
Protuberance, surface folding is opaque, and bacterium colony is white (Fig. 2A).The bacterial strain B-11 observed under transmission electron microscope is in the shape of a rod or cylinder
Shape, the more flagellums of Zhousheng, size are (0.94~1.18) μ m (2.93~3.94) μm (Fig. 2 B).Its feature and bacillus
(Bacillus sp.) is consistent.
Reference《Common bacteria identification handbook》The physio-biochemical characteristics of the culture based assays bacterial strain ZJU-C612-2 of middle recommendation
It was found that oxidizing ferment, catalase, V-P measurement, Starch Hydrolysis, lecithinase, indoles experiment are the positive, methyl red is feminine gender
(table 1).Reference《Common bacteria system identification handbook》, lecithin cannot be generated according to bacillus subtilis and bacillus amyloliquefaciens
Esterase determines that bacterial strain ZJU-C612-2 is Bei Laisi bacillus.
The physio-biochemical characteristics of table 1, bacterial strain ZJU-C612-2
Table 1Physiological and biochemical characteristics of strain ZJU-
C612-2
Embodiment 3, bacterial strain ZJU-C612-2 produce the measurement of antibacterial substance
1, the measurement of protease ability is produced
Bacterial strain ZJU-C612-2 point is connected to protease detection plate (A：Skimmed milk power 8g is dissolved in 300mL distilled water, and 115
DEG C sterilizing 10min, B：Agar 8g adds distilled water to be settled to 300mL, and 121 DEG C of sterilizing 20min, A are mixed after being sterilized separately with B) on,
30 DEG C of dark culture 2d, the discovery edge bacterial strain ZJU-C612-2 generate apparent transparent circle (Fig. 3 A), illustrate bacterial strain ZJU-C612-2
Protease can be generated.
2, the measurement of chitinase ability is produced
Bacterial strain ZJU-C612-2 point is connected to the culture medium (ammonium dihydrogen phosphate using colloidal chitin as sole carbon source
1.0g, potassium chloride 0.2g, Magnesium sulfate heptahydrate 0.2g, w=1% colloidal chitin 100mL, agar 20g, distilled water 1000mL,
PH7.0), 30 DEG C of culture 3d, discovery bacterial strain ZJU-C612-2 do not have transparent circle (Fig. 3 B), illustrate that bacterial strain ZJU-C612-2 can not
Generate chitinase.
3, the measurement of 1,4 beta-glucanase ability is produced
Bacterial strain ZJU-C612-2 point is connected to aniline blue glucan agar medium plate (glucose 1.0g, Poria cocos powder
4.0g, dipotassium hydrogen phosphate 1.0g, disodium hydrogen phosphate 3.0g, aniline blue 0.06g, green vitriol 0.5g, agar powder 12g,
Distilled water 1000mL, pH 7.0), 30 DEG C of 3~5d of culture, discovery bacterial strain ZJU-C612-2 has transparent circle (Fig. 3 C), illustrates bacterial strain
ZJU-C612-2 can generate 1,4 beta-glucanase.
4, thermophilic iron element ability measurement is produced
Bacterial strain ZJU-C612-2 point, which is connected to chromium Austria alcohol (CAS) culture medium, (takes 60.5mg chromium Austria alcohol to be dissolved in 50mL Fe3+
(1mmol/L FeCl3·6H2O, 10mmol/L HCl) solution be uniformly mixed, while agitating, be slowly added 16 amino
After baked base ammonium bromide (HDTMA) solution (72.9mg is dissolved in 40mL water), pH to 7.0 is adjusted, 100mL is then settled to.It finally will be upper
State after solution sterilization with 900mL WA culture medium (agar containing 20g, peptone 4.5g, glucose 9g, beef extract 2.7g, NaCl
4.5g, pH 7.0) mixing), 30 DEG C of 3~5d of culture, discovery bacterial strain ZJU-C612-2 has crocus haloing (Fig. 3 D), illustrates bacterial strain
ZJU-C612-2 can generate thermophilic iron element.
5, the measurement of lipopeptide antibiotic ability is produced
Bacterial strain ZJU-C612-2 is lined 37 DEG C of constant temperature incubations on LB solid medium, and for 24 hours, each bacterial strain after activation is pressed
5% volume ratio is inoculated in LB liquid medium, and 37 DEG C, 200rpm, shaken cultivation for 24 hours, as primary seed solution, then presses 5%
Volume ratio is inoculated in Landy culture medium, and secondary seed solution is cultivated under similarity condition, and the latter is coated on after being diluted with sterile water
37 DEG C of constant temperature incubation 48h of Landy solid medium.2 single bacteriums of picking are fallen on the target of Target Board hole, add 1 μ L Extraction solvent, 1 μ L
Auxiliary matrix mixes, and MALDI-TOF-MS is detected after natural air drying.MALDI-TOF-MS testing result shows bacterial strain ZJU-C612-
2 can generate withered grass plain (Iturin), Feng Yuansu (Fengycin) and Surfactin (Surfactin) three classes lipopeptid class antibiosis
Plain (table 2, Fig. 4).
Her withered grass element, Feng Yuansu, Surfactin testing result of 2 bacterial strain ZJU-C612-2 of table
The measurement of 4 bacterial strain ZJU-C612-2 of embodiment production plant hormone
The fermentation liquid (fermentation condition is with embodiment 1) of 1L bacterial strain ZJU-C612-2 is taken, 5000rpm is centrifuged 5min, is trained
Nutrient solution is separately added into isometric ethyl acetate in culture solution, after being sufficiently stirred, stands extraction until removing after layering organic
Phase, and extracted (extraction mode is same as above) again to water phase, continuous 3 collections merge organic solvent phase, with Rotary Evaporators into
Row is evaporated under reduced pressure, and obtains enriched product, is that 20mL methanol dissolves enriched product with total volume, obtains hormone extracting solution,
Finally with the content of high performance liquid chromatography-mass spectrometry instrument (LC-MS) measurement GA, KT, ZT, IAA and SL, measurement result display hair
It is 1.2ng/L that the content of its GA, which is the content that the content of 0.22 μ g/L, KT is 0.10 μ g/L, ZT, in zymotic fluid, and the content of IAA is
The content of 31.31 μ g/L, SL is 0.50 μ g/L (Fig. 5).
Embodiment 5, bacterial strain ZJU-C612-2 are to the bacteriostatic experiments of 6 kinds of plant pathogens
Take the dendrobium candidum anthrax bacteria, Crocus bulb rotten pathogenic bacteria, walnut branch-rot bacterium, rape sclerotium of diameter 5mm
6 kinds of germ, cotton standing dead silk core germ and Didymella bryoniae plant pathogen bacteria cakes are respectively placed in the left side distance of PDA plate
At central 20mm, the aseptic filter paper piece of diameter 5mm is correspondingly placed on the right side of the center at 20mm and accesses biocontrol bacteria ZJU-
5 μ L of C612-2 bacterium solution (fermentation condition is with embodiment 1) is control to be inoculated with the 5 sterile LB of μ L, and every processing is repeated 3 times.28 DEG C of constant temperature
Culture, measurement pathogenic bacteria stop measurement after increment tie-in 3d is unchanged, use ruler close to the mycelium morphology factor of endophyte side
Measure the antibacterial bandwidth between biocontrol microorganisms and pathogenic bacteria.Measurement result shows that bacterial strain ZJU-C612-2 is able to suppress dendrobium candidum charcoal
Subcutaneous ulcer germ, Crocus bulb rotten pathogenic bacteria, walnut branch-rot bacterium, Sclerotinia sclerotiorum, cotton standing dead silk core germ and watermelon are climing
The plant pathogen of 6 kind of blight bacterium, antibacterial bandwidth be followed successively by 19.5 ± 0.1mm, 11.2 ± 0.5mm, 5.7 ± 1.2mm,
17.0 ± 0.8mm, 16.5 ± 0.7mm, 13.8 ± 1.4mm (Fig. 6).
Embodiment 6, bacterial strain ZJU-C612-2 are to the Biological control of dendrobium candidum protocorm and tissue-cultured seedling
Cultivating minimal medium formula used in dendrobium candidum protocorm and tissue-cultured seedling culture is：MS+BA 0.5mg/L+
NAA0.1mg/L+ potato 50g+ agar 7.5g+ sucrose 30g, pH=5.5~5.8, this culture medium can guarantee dendrobium candidum protocorm
The primary demand of stem and tissue-cultured seedling growth.Culture dendrobium candidum protocorm and the condition of tissue-cultured seedling are：16h illumination, intensity of illumination
For 30~40 μm of olm-2·s-1, temperature is (27 ± 1) DEG C；8h dark culture, temperature are (21 ± 1) DEG C；Above-mentioned illumination and dark
Alternately.
Experimental comparison group is the sterile LB of addition 3% in minimal medium, and processing group is addition 3% in minimal medium
LB shakes the bacterial strain ZJU-C612-2 fermentation liquid of training (fermentation condition is with embodiment 1).The selection of protocorm Biological control is inoculated on the same day
Homogeneous, color similar in protocorm be inoculated with, be inoculated with 0.5g protocorm, each processing in when inoculation each tissue culture bottle
15 bottles of inoculation, is repeated 3 times, and weighs again after cultivating 45d, calculates protocorm and increases weight and increased percentage.Tissue-cultured seedling growth-promoting
Experiment selects the inoculation uniform candidum tissue culturing seedling of growing way (plant height 1cm or so) to be on the same day inoculated with, and each tissue culture bottle connects
10, when inoculation, cuts off root, is close to bottle wall and is inoculated with, and is saved after inoculation with the plant height record that ruler measures every tissue-cultured seedling
And number, each processing is inoculated with tissue-cultured seedling 60, is repeated 3 times, and measures plant height again after cultivating 45d, calculates increased percentage, together
Shi Tongji lateral bud quantity.It is shown the experimental results showed that bacterial strain ZJU-C612-2 fermentation liquid has the growth of dendrobium candidum protocorm
Facilitation is write, the protocorm through bacterial strain ZJU-C612-2 fermentation liquor treatment increases weight and improve 35% compared with sterile LB, knot
Fruit is shown in Table 3, Fig. 7；Bacterial strain ZJU-C612-2 fermentation liquid also has the effect of remarkably promoting to the growth of candidum tissue culturing seedling, through bacterium
The protocorm of strain ZJU-C612-2 fermentation liquor treatment increases height and improves 50%, lateral bud number improves 28%, knot compared with sterile LB
Fruit is shown in Table 4, Fig. 8.
Table 3, bacterial strain ZJU-C612-2 are to the growth-promoting effect of dendrobium candidum protocorm
Growth-promoting effect of the 4 bacterial strain ZJU-C612-2 of table to iron sheet dry measure used in former times tissue-cultured seedling
Existing Bei Laisi bacillus is carried out bacteriostatic experiment, acquired results by comparative experiments 1 as described in embodiment 5
Comparison with invention bacterial strain ZJU-C612-2 is described in table 5 below.
Table 5, antibacterial bandwidth
| Walnut branch-rot bacterium | Didymella bryoniae | |
| Bei Laisi bacillus velezensis ZJU-C612-2 | 5.7±1.2mm | 13.8±1.4mm |
| Bei Laisi bacillus (CP023414.1) | 2~3mm | 5~7mm |
| Bei Laisi bacillus (Bacillus velezensis) bacterial strain ZJ20 | 2~3mm | 4~6mm |
| Bei Laisi bacillus (Bacillus velezensis) CGMCC No.4992 | 3~4mm | 5~8mm |
| Bei Laisi bacillus CC09 (Cai et al., 2017) | 1~2mm | 4~6mm |
| Bei Laisi bacillus DL-59 (Shuna et al., 2011) | 2~4mm | 5~8mm |
| Bei Laisi bacillus RC218 (Juan et al., 2007) | 3~4mm | 4~6mm |
| Bei Laisi bacillus BAC03 | 1~2mm | 3~5mm |
Comparative experiments 2, existing Bei Laisi bacillus is carried out as described in embodiment 6 to dendrobium candidum protocorm and
The comparison of the Biological control of tissue-cultured seedling, acquired results and invention bacterial strain ZJU-C612-2 is described in table 6 below.
Table 6
The above list is only a few specific embodiments of the present invention for finally, it should also be noted that.Obviously, this hair
Bright to be not limited to above embodiments, acceptable there are many deformations.Those skilled in the art can be from present disclosure
All deformations for directly exporting or associating, are considered as protection scope of the present invention.
Claims (3)
1. Bei Laisi bacillus velezensis ZJU-C612-2, it is characterized in that：For Bei Laisi bacillus
ZJU-C612-2Bacillus velezensis ZJU-C612-2, deposit number：CCTCC NO：M 2018312.
2. the purposes of Bei Laisi bacillus velezensis ZJU-C612-2 as described in claim 1,
It is characterized in：Inhibit dendrobium candidum anthrax bacteria (Colletotrichum gloeosporioides), Crocus bulb rot disease
Bacterium (Fusarium oxysporum), walnut branch-rot bacterium (Botryosphaeria dothidea), Sclerotinia sclerotiorum
(Sclerotinia sclerotiorum), cotton standing dead silk core germ (Rhizoctonia solani), Didymella bryoniae
The growth of (Mycosphaerella melonis).
3. the purposes of Bei Laisi bacillus velezensis ZJU-C612-2 as described in claim 1,
It is characterized in：Promote dendrobium candidum protocorm and tissue-cultured seedling growth.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109554317A (en) * | 2018-12-29 | 2019-04-02 | 陕西省微生物研究所 | Bei Laisi bacillus and its application in wheat sharp eyespot disease prevention growth-promoting |
| CN109880764A (en) * | 2019-03-12 | 2019-06-14 | 河北农业大学 | One plant of Bei Laisi bacillus and its application in prevention and treatment apple disease |
| CN110301335A (en) * | 2019-07-10 | 2019-10-08 | 浙江大学 | The cultivation of the lateral bud breeding bulb of west safflower |
| CN111019866A (en) * | 2019-12-30 | 2020-04-17 | 华南理工大学 | Endophytic bacillus of Pu' er tea tree leaves and application thereof |
| CN111484946A (en) * | 2019-01-28 | 2020-08-04 | 福建省农业科学院农业生物资源研究所 | IAA-producing high-temperature-resistant bacillus and application thereof |
| WO2023142162A1 (en) * | 2022-01-26 | 2023-08-03 | 杭州师范大学 | Bacillus strain wyj-e14 isolated from curcuma wenyujin y. h. chen & c. ling and use thereof in preparation of anti-tumor drug |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108102991A (en) * | 2018-03-07 | 2018-06-01 | 山东戴盟得生物科技有限公司 | One plant of Bei Laisi bacillus and its application in watermelon blight is prevented |
-
2018
- 2018-06-27 CN CN201810682341.XA patent/CN108865930B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108102991A (en) * | 2018-03-07 | 2018-06-01 | 山东戴盟得生物科技有限公司 | One plant of Bei Laisi bacillus and its application in watermelon blight is prevented |
Non-Patent Citations (3)
| Title |
|---|
| XUN-CHAO CAI等: "Genomic and metabolic traits endow Bacillus velezensis CC09 with apotential biocontrol agent in control of wheat powdery mildew disease", 《MICROBIOLOGICAL RESEARCH》 * |
| 张莹等: "生防芽孢杆菌分泌的拮抗物质的研究进展", 《浙江农业科学》 * |
| 徐桑尔等: "多粘类芽孢杆菌防治铁皮石斛细菌性叶斑病及促生效果", 《浙江农业科学》 * |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109554317A (en) * | 2018-12-29 | 2019-04-02 | 陕西省微生物研究所 | Bei Laisi bacillus and its application in wheat sharp eyespot disease prevention growth-promoting |
| CN111484946A (en) * | 2019-01-28 | 2020-08-04 | 福建省农业科学院农业生物资源研究所 | IAA-producing high-temperature-resistant bacillus and application thereof |
| CN111484946B (en) * | 2019-01-28 | 2022-08-19 | 福建省农业科学院农业生物资源研究所 | IAA-producing high-temperature-resistant bacillus and application thereof |
| CN109880764A (en) * | 2019-03-12 | 2019-06-14 | 河北农业大学 | One plant of Bei Laisi bacillus and its application in prevention and treatment apple disease |
| CN109880764B (en) * | 2019-03-12 | 2021-10-26 | 河北农业大学 | Bacillus belgii and application thereof in prevention and treatment of apple diseases |
| CN110301335A (en) * | 2019-07-10 | 2019-10-08 | 浙江大学 | The cultivation of the lateral bud breeding bulb of west safflower |
| CN110301335B (en) * | 2019-07-10 | 2021-10-01 | 浙江大学 | Culture method for lateral bud breeding corm of saffron |
| CN111019866A (en) * | 2019-12-30 | 2020-04-17 | 华南理工大学 | Endophytic bacillus of Pu' er tea tree leaves and application thereof |
| CN111019866B (en) * | 2019-12-30 | 2022-08-16 | 华南理工大学 | Endophytic bacillus of Pu' er tea tree leaves and application thereof |
| WO2023142162A1 (en) * | 2022-01-26 | 2023-08-03 | 杭州师范大学 | Bacillus strain wyj-e14 isolated from curcuma wenyujin y. h. chen & c. ling and use thereof in preparation of anti-tumor drug |
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